[1]刘亚军,闫峰,叶巍,等.转化生长因子-β2（TGF-β2）诱导人视网膜色素上皮层细胞上皮-间质转分化中miRNA-29b的表达及意义[J].眼科新进展,2016,36(11):1001-1005.[doi:10.13389/j.cnki.rao.2016.0268]
　LIU Ya-Jun,YAN Feng,YE Wei,et al.Expression changes and significance of miRNA-29b in TGF-β2 induced epithelial mesenchymal transition of human retinal pigment epithelial cells[J].Recent Advances in Ophthalmology,2016,36(11):1001-1005.[doi:10.13389/j.cnki.rao.2016.0268]
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转化生长因子-β2（TGF-β2）诱导人视网膜色素上皮层细胞上皮-间质转分化中miRNA-29b的表达及意义
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《眼科新进展》[ISSN:1003-5141/CN:41-1105/R]

卷:
36卷
期数:
2016年11期
页码:
1001-1005
栏目:
实验研究
出版日期:
2016-11-05

文章信息/Info

Title:
Expression changes and significance of miRNA-29b in TGF-β2 induced epithelial mesenchymal transition of human retinal pigment epithelial cells
作者:
刘亚军; 闫峰; 叶巍; 朱欣悦; 黄振平
210002　江苏省南京市，南京大学医学院临床学院，南京军区南京总医院眼科（刘亚军，朱欣悦）；210002　江苏省南京市，南京军区南京总医院眼科（闫峰，黄振平）；200082　上海市，第二军医大学南京军区南京总医院眼科（叶巍）
Author(s):
LIU Ya-Jun; YAN Feng ; YE Wei; ZHU Xin-Yue;  HUANG Zhen-Ping
Department of Ophthalmology,School of Medicine,Nanjing University,Nanjing General Hospital of Nanjing Military Command of PLA(LIUYa-Jun,ZHUXin-Yue),Nanjing210002,Jiangsu Province,China;Department of Ophthalmology,Nanjing General Hospital of Nanjing Military Command of PLA(YANFeng,HUANGZhen-Ping),Nanjing210002,Jiangsu Province.China;Department of Ophthalmology,School of the Second Mititary Medical University,Nanjing Ceneral Hospital of Nanjing Military Command of PLA(YEWei),Shanghai200082,China
关键词:
人视网膜色素上皮层细胞; 上皮-间质转分化; 转化生长因子-β2; miRNA-29b; 增生性玻璃体视网膜病变
Keywords:
human retinal pigment epithelial cells ;  epithelial mesenchymal transition ;  transforming growth factor β2 ;  miRNA-29b ;  proliferative vitreoretinopathy
分类号:
Ｒ７７４.１
DOI:
10.13389/j.cnki.rao.2016.0268
文献标志码:
A
摘要:
目的　探究转化生长因子-β２（ｔｒａｎｓｆｏｒｍｉｎｇｇｒｏｗｔｈｆａｃｔｏｒ-β２，ＴＧＦ-β２）诱导人视网膜色素上皮（ｒｅｔｉｎａｌｐｉｇｍｅｎｔｅｐｉｔｈｅｌｉａｌ，ＲＰＥ）层细胞上皮-间质转分化中ｍｉＲＮＡ-２９ｂ的表达变化及意义。方法　使用不同浓度ＴＧＦ-β２刺激ＲＰＥ细胞上皮-间质转分化后，倒置相差显微镜观察细胞形态变化，Ｗｅｓｔｅｒｎｂｌｏｔ、ＲＴ-ＰＣＲ检测成纤维化相关分子纤维连接蛋白（ｆｉｂｒｏｎｅｃｔｉｏｎ，ＦＮ）、神经钙粘连蛋白（ｎｅｒｖｅｃａｌｃｉｕｍａｄｈｅｓｉｏｎｐｒｏ-ｔｅｉｎ，Ｎ-Ｃａｄｈｅｒｉｎ）的表达。采用ＲＴ-ＰＣＲ检测不同浓度ＴＧＦ-β２及不同时间ＴＧＦ-β２（５μｇ·Ｌ－１）刺激ＲＰＥ细胞后ｍｉＲＮＡ-２９ｂ的表达。结果　ＴＧＦ-β２刺激后的ＲＰＥ细胞形态呈纤维化改变。在一定浓度范围内（１μｇ·Ｌ－１、５μｇ·Ｌ－１、１０μｇ·Ｌ－１），随着ＴＧＦ-β２浓度的增加ＦＮ、Ｎ-Ｃａｄｈｅｒｉｎ及相应ｍＲＮＡ随之增加，１μｇ·Ｌ－１、５μｇ·Ｌ－１、１０μｇ·Ｌ－１组与对照组（０μｇ·Ｌ－１）相比，差异均有统计学意义（均为Ｐ＜０．０１）。ＴＧＦ-β２在一定浓度范围内（０μｇ·Ｌ－１、１μｇ·Ｌ－１、５μｇ·Ｌ－１）以剂量依赖的方式诱导ＲＰＥ细胞ｍｉＲＮＡ-２９ｂ表达的降低，１μｇ·Ｌ－１、５μｇ·Ｌ－１、１０μｇ·Ｌ－１组与对照组（０μｇ·Ｌ－１）相比，差异均有统计学意义（均为Ｐ＜０．０１），在ＴＧＦ-β２浓度为５μｇ·Ｌ－１时ｍｉＲＮＡ-２９ｂ表达量最低。ＴＧＦ-β２在一定时间范围内（０ｈ、３ｈ、６ｈ、１２ｈ）以时间依赖的方式诱导ＲＰＥ细胞ｍｉＲＮＡ-２９ｂ表达的降低，３ｈ、６ｈ、１２ｈ、２４ｈ、４８ｈ组与０ｈ相比差异均有统计学意义（均为Ｐ＜０．０５）。结论　一定范围内ＴＧＦ-β２以剂量和时间依赖的方式诱导ＲＰＥ细胞ｍｉＲＮＡ-２９ｂ的表达，为进一步研究ｍｉＲＮＡ-２９ｂ与ＴＧＦ-β２在ＲＰＥ细胞上皮-间质转分化过程中的相互作用提供理论基础。
Abstract:
Objective To explore the expression changes and significance of miRNA-29b in TGF-[32 induced epithelial mesenchymal transition ( EMT) of human retinal pigment epithelial ( RPE ) cells. Methods Different concentrations of TGF-[32 (0 vg . 1’1 ,1 Vg . 1’] ,5 Vg . L",10 Vg . L") were used to stimulate EMT of RPE cells . and then the changes of cell morphology were observed by inverted phase contrast microscope. The expression of fibronectin ( FN) and nerve calcium adhesion protein ( N-Cadherin) were detected by Western blot and RT-PCR. RT-PCR was used to detect the expression of miRNA-29b in RPE cells after stimulated with TGF-[32 at different concentrations(0 Vg . 1’1 ,1 Vg . 1-1 ,5 Vg ’ L -l , 10 Vg . L -l ) and at different time points (0 hour , 3 hours .6 hours . 12 hours . 24 hours.48 hours) of TGF-[3z(5 Vg . L"). Results RPE cells exhibited a fiber-like morphology after stimulated with TGF-[32. With the increase of TGF-[32 concentration (1 yg . 1’1 ,5 Vg . 1’1 ,10 Vg . L") and the expression of FN, NCadherin protein and the corresponding mRNA increased. Other groups ( I Vg ’ L-’,5 Vg . L",10 Vg . L-’) compared with blank control group (0 vg . L-’) , the differences were significant ( all P < 0. 01). RPE cells induced by TGF-[32 showed a decreasing expression of nuRNA-29b in a dose dependent manner (0 vg . L",1 Vg ’ L-’,5 Vg ’ L"). Other groups (1 Vg ’ L",5 Vg ’ L-’, 10 Vg . L") compared with blank control group (0 Vg . L-’) and the differences were significant ( all P < 0. 01 ). When the TGF-[32 concentration was 5 yg . L-l , the expression of miRNA-29b reached the lowest. RPE cells induced by TGF-[32 showed a decreasing expression of miRNA-29b in a time dependent manner (0 hour , 3 hours . 6 hours , 12 hours ) . Other groups ( 3 hours . 6 hours . 12 hours . 24 hours.48 hours) compared with blank control group (0 hour) and the differences were sigruficant ( all P < 0. 05 ) . Conclusion TGF- [32 can induce the expression of miRNA-29b in RPE cells in a time and dose-dependent manner in a certain range,which provide a theoretical basis for the further study of the cross-talk nuRNA-29b and TGF-[32 in the EMT process of RPE cells.

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[2]丁芝祥,陈阳,秦贤杰,等.miRNA-184在TGF-β2诱导人晶状体上皮细胞上皮-间质转分化中的表达[J].眼科新进展,2015,35(12):1116.[doi:10.13389/j.cnki.rao.2015.0306]
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备注/Memo

备注/Memo:
国家自然科学基金面上项目（编号：８１２７０９７９）
更新日期/Last Update: 2016-11-29